SBIR Phase I: New approach for producing increased quantities of human antibodies for research, medical, and biodefense applications

该小型企业创新研究（SBIR）项目的更广泛影响/商业潜力将满足抗体市场对快速廉价生产抗体的需求。抗体是生命科学中最常用的分子工具之一，其应用跨越生物科学和医学的各个领域。2008年全球抗体市场估计约为400亿美元，预计到2016年将超过680亿美元。然而，开发新的抗体是一个漫长而昂贵的过程。 我们的目标是开发一种系统，大大减少开发新抗体的时间和成本。 所提出的系统和所得抗体可用作未来治疗剂的来源，用作生物防御和医学中的检测系统的组分，或用作研究和诊断的亲和纯化系统。 该系统将在第一阶段进行展示，生产针对肉毒杆菌神经毒素的抗体，这将对生物防御领域具有价值。该SBIR项目旨在开发一种快速生产针对特定应用的大量抗体的系统。第一阶段的主要目标是开发一个平台，该平台允许稳健地产生产生大量抗体的克隆，而不需要耗时和劳动密集型的基因扩增技术，如脱氢叶酸还原酶或谷氨酰胺合酶选择系统。这将与先前开发的方法相结合，该方法允许同时生成多种人源化或完全人源化抗体，以快速生成可以满足抗体市场每个细分市场中存在的特定需求的各种抗体集合。 然而，通过将构建体直接转染到次级细胞系（如中国卵巢（CHO）和人胚肾（HEK））中产生的克隆产生的抗体的量显著低于常规杂交瘤。生产的抗体量不足以满足抗体市场任何部分的需求。为了解决抗体市场所有领域中存在的需求，提出了开发一种新的方法，用于快速有效地产生和选择免疫球蛋白产生水平将与平均杂交瘤的免疫球蛋白产生水平相当的细胞克隆。与用于免疫球蛋白编码构建体的稳健组装的系统组合，该方法将允许快速且稳健地产生适用于特定应用的抗体。

The broader impact /commercial potential of this Small Business Innovation Research (SBIR) project will address the needs of the antibody market for fast and inexpensive production of antibodies. Antibodies are among the most utilized molecular tools in the life sciences, whose applications span a variety of areas in biological science and medicine. The worldwide antibody market was estimated at approximately $40 billion in 2008, and is expected to rise above $68 billion by 2016. The development of new antibodies, however, is a lengthy and expensive process. The goal is to develop a system that significantly reduces the time to develop new antibodies as well as the cost. The proposed system and resulting antibodies may be used as a source of future therapeutics, as components of detection systems in biodefense and medicine, or as affinity purification systems for research and diagnostics. The system will be demonstrated in Phase I with the production of antibodies targeted at botulinum neurotoxin, which will be of value to the biodefense area.This SBIR project aims to develop a system for the fast generation of high quantities of antibodies targeted at specific applications. The main objective in Phase I is to develop a platform that allows robust generation of clones producing high quantities of antibodies without the need for time-consuming and labor-intensive gene amplification techniques, such as the dehydrofolate reductase or the glutamine synthase selection systems. This will be coupled with a previously developed method that allows simultaneous generation of multiple humanized or completely human antibodies for the fast generation of diverse collections of antibodies that can address specific needs existing in each segment of the antibody market. However, clones generated via direct transfection of constructs into secondary cell lines, such as Chinese Hamster Ovary (CHO) and Human Embryo Kidney (HEK), produce substantially lower amounts of antibodies than do conventional hybridomas. The amount of antibodies produced is not sufficient to address the needs in any segment of the antibody market. To address needs existing in all segments of the antibody market, it is proposed to develop a new approach for fast and efficient generation and selection of cell clones whose immunoglobulin production levels will be comparable to those of average hybridomas. In combination with the system for robust assembly of immunoglobulin-encoding constructs, this approach will allow the rapid and robust generation of antibodies adapted for use in specific applications.